Varietal discrimination and genetic relationships of Vitis vinifera L. cultivars from two major Controlled Appellation (DOC) regions in Portugal

Thirty-nine grapevine cultivars widely grown in Portugal, especially in Vinhos Verdes and Douro regions, and two well known international cultivars as standards, were genotyped at 12 microsatellite loci. The number of alleles per locus ranged from 6 to 12, and the number of allelic combinations per locus from 13 to 26. The total number of unique genotypes in the 12 analysed loci was 120, having most of the cultivars (38 out of 41) at least one unique genotype in any of the loci. The microsatellite profiles were adequate to discriminate 41 cultivars. The level of observed heterozygosity at each locus varied from 70.7% to 95.1%. VVMD28 has been revealed as one of the most informative markers. Several synonymies between Spanish and Portuguese cultivars were confirmed, and some homonymies are discussed. The genetic profiles of all 41 cultivars were searched for possible parent-offspring groups. The data obtained revealed the possible descendence of Touriga Franca from Touriga Nacional and Marufo.     

Microsatellite fingerprinting of homonymous grapevine (Vitis vinifera L.) varieties in neighboring regions of South-East Turkey

Genotyping of Turkish grapevine (Vitis vinifera L.) germplasm was characterized by use of six highly polymorphic microsatellite loci (VVS2, VVMD5, VVMD7, VVMD27, VrZAG62, VrZAG79). In this study we aimed to clarify the relationships between homonymous varieties coming from different regions. Our results showed a large degree of genetic variability among most of the homonymous cultivars. The number of alleles per locus ranged from 10 to 21, and gene diversity (expected heterozygosity) values ranged from 0.85 to 0.93. Cultivars presenting the same names of Sergi karas? (sampled from ?anl?urfa and Gaziantep), Yediveren (sampled from ?anl?urfa, Gaziantep, and National Germplasm Repository Vineyard in Tekirda?) and Serpenek?ran (sampled from ?anl?urfa and Gaziantep) were clustered together, or very close to each other, in a phenogram. Moreover, the alleles at the six microsatellite loci analyzed were found to be similar in terms of base pairs within each of these three closely positioned varieties. However, all the other cultivars failed to show a suitable clustering pattern when comparing their DNA profiles and names. Similarly named cultivars were not generally grouped together in the phenogram. On the other hand, we detected a tendency for differently named homonymous grape cultivars to cluster together.     

Use of chloroplast microsatellite markers as a tool to elucidate polymorphism,classification and origin of Tunisian grapevines

Chloroplast microsatellite markers were used in this study to genotype 43 grapevines accessions grown in Tunisia. Size variation was observed for the three cpSSR loci, both in the sample of cultivars and in wild accessions. The seven alleles observed in the sample of cultivars for the three loci are present in wild accessions except that their distribution is different. Levels of genetic diversity obtained for the Tunisian grapevines either in wild or cultivated gene pools are high and comparable with values obtained with other studied samples of Vitis vinifera. The distribution of haplotypes within the two samples is differential. Indeed, the chlorotype A is most abundant in the wild sample, whereas the chlorotype C is majority in the sample of cultivars. Haplotypes frequencies for cultivated grapevine distinguish haplotypes B and C as the most frequent (28% and 44% respectively) and haplotypes A and D as the least frequent (16% and 12% respectively). For wild grapevines, the seven alleles combined in three haplotypes, A, C and D. The haplotype A is the most frequent (44%) in the analyzed sample of wild accessions while haplotypes C and D show a frequency of 28%. Chlorotype distribution in Tunisian cultivars is comparable with that of cultivars in the Eastern Region representing the primary centre of domestication of the species. These results agree with the higher relevance of table grape cultivars in Tunisian viticulture and support an oriental origin of a large part of autochthons cultivars. Our results agree with other studies based in nuclear and chloroplast microsatellite markers and suggest independent domestication events for V. vinifera L. species.     

Ampelographic and genetic characterisation of ancestral grapevine (Vitis vinifera L.) accessions present in the Umbria Region (Central Italy)

Summary

During an ongoing effort to recover and preserve local germplasm, 14 accessions of indigenous minor grapevine (Vitis vinifera L.) cultivars from the Umbria Region, Central Italy, were chosen because they had been neglected and were threatened with extinction. Their phenotypic and genetic characteristics were evaluated through an ampelographic study of their shoots, mature leaves, bunches, and berries and by genomic analysis using an international set of nine microsatellite (simple sequence repeat; SSR) markers (VVS2, VVMD5, VVMD7, VVMD27, VrZAG62, VrZAG79, VVMD25, VVMD28, and VVMD32). Comparisons of the SSR profiles of all 14 accessions with grapevine accessions in several databases permitted the identification of unique genotypes, as well as possible synonyms. Information on these older, neglected cultivars will help to reduce the genetic erosion of grapevine germplasm, improve conservation and possible recovery, and assist in the future production of new, distinctive wines.     

Characterization for yield, fruit quality, and molecular profiles of lemon genotypes tolerant to ‘mal secco’ disease

‘Mal secco’ is a serious disease of lemon and other citrus species, reduces lemon production, and limits the use of susceptible cultivars especially in the Mediterranean countries, against which there is no effective management system to control the disease. The objective of this study was to determine yield efficiency, tree and fruit characters, and molecular profiles of the selected promising genotypes in lemon breeding program to develop ‘mal secco’ tolerant lemon cultivars. After 4 years of evaluation, ‘Tuzcu 894’, a progeny of ‘Finike Yerli Yuvarlak’ and ‘Kutdiken’, was designated as promising genotype with high yield, fruit weight, juice content and low seed number contrary to ‘Kutdiken’ as control. ‘Tuzcu 8911’ was suitable for dense planting having the highest yield in CV. In addition, these two promising genotypes survived after many years being subjected to artificial and natural P. tracheiphila infections and harsh winter temperatures, which were attractive to the citrus industry. Molecular analysis, as assessed with 11 inter-simple sequence repeats (ISSR) primers, indicated that ‘Tuzcu 09 Aklimon’ and its selections, ‘Tuzcu 896’, ‘Tuzcu 897’ and ‘Tuzcu 898’, were distinct from the other genotypes with similarity value of 0.82. Genetic variation among the other lemon genotypes was low and some of the genotypes were identical. It was concluded that variations in the agronomical characters are mainly due to mutations.     

Discrimination of the grapevine cultivars ‘Picolit’ and ‘Kéknyelű’ with molecular markers

The name of the grapevine cultivar ‘Kéknyel?’ has become inseparable from the name of the Badacsony vine region, whose fame is well known beyond our frontier as well. In the Vitis International Variety Catalogue (http://www.genres.de/idb/vitis/) ‘Kéknyel?’ is reported, as the synonym of the Italian grapevine cultivar ‘Picolit’. Vertical poliacrylamide-gel electrophoresis was used for the investigation isoenzymes of catechol-oxidase (CO) and acid phosphatase (AcP). Microsatellite analyses were carried out at 6 loci (VVS2, VVS16, VrZag79, VVMD7, VMC4A1, VMC4G6). The results of the isoenzymatic and microsatellite analyses confirmed, that this two cultivars are different.     

Immunolocalization and quantification of IAA after self- and free-pollination in Olea europaea L.

In order to characterise the self-compatibility in ‘Pisa 9’, that is an improved clone of cultivar Leccino (Olea europaea L.), biological, histological and biochemical analyses were carried out. Observations were also focused on ‘Pendolino’, the main pollinator for Italian cultivars considered as being self-incompatible. After self-pollination, in ‘Pisa 9’ the majority of the pollen tubes reached the distal part of the style, entered the ovule leading to a high fruit-set percentage. These same processes were also noted in the Pendolino cultivar, though in a lesser degree. The indole acetic acid (IAA) concentration and its immunocytochemical localization were determined in pollinated flowers after self- and free-pollination and in unpollinated flowers. In both cultivars the highest IAA levels and signal intensity were observed in free-pollinated pistils, whereas after self-pollination a different behaviour emerged. In ‘Pendolino’ the immunostaining was not uniformly distributed and stopped at the style level. This finding sustained the low percentage of fruit-set obtained in the cultivar. In ‘Pisa 9’ the auxin distribution was similar to the free-pollinated pistils where the immunolocalization staining reached the ovary. The correspondence between the IAA signal in the ovary and the high percentage of fruit-set in ‘Pisa 9’ self-pollinated pistils, confirms the role of this hormone in the induction and regulation of the fertilisation process.     

Study of long-distance movement of Plum pox virus (Sharka) as an alternative resistance-evaluation method in Prunus

During the breeding programs for Plum pox virus (PPV, Sharka) resistance in Prunus, the evaluation of the new releases through symptoms observation on leaves has been contradictory and represents one of the main handicaps in these programs. In order to increase the accuracy of this traditional evaluation method, we here analyze an alternative method based on the study of the ability of a genotype to allow the long-distance movement of the virus through its vascular vessels. Two different plant models have been assayed: (a) in Model I, the inoculation was performed in the ‘GF305’ rootstock with a later grafting of the genotype under evaluation and a scion of healthy control ‘GF305’, to evaluate the long-distance movement through the studied genotype from the rootstock to the scion (xylem transport), and (b) in Model II, the inoculation with ‘GF305’ diseased scions was performed by grafting these diseased scions onto the studied genotypes, which were grafted onto healthy ‘GF305’ peach seedlings, to evaluate the long-distance movement through the studied genotype from the scion to the rootstock (phloem transport). The results show that, regardless of the presence of symptoms, susceptible genotypes allowed the movement of the virus through their vascular vessels in both directions studied. However, the resistant apricot ‘Stark Early Orange’ did not allow this movement. We propose the study of the ability of a genotype to allow the long-distance movement of the virus as an alternative and more accurate method for the evaluation of PPV resistance. However, this protocol is much more tedious than the traditional one and could be used mainly in the evaluation of a reduced number of more interesting genotypes.     

Morphogenic gradients of adventitious bud and shoot regeneration in epicotyl explants of Citrus

The in vitro responses of epicotyl explants from ‘Cravo’ rangpur lime (Citrus limonia Osb.), ‘Foster’ grapefruit (C. paradisi Macf.), and ‘Pera’ sweet orange (C. sinensis (L.) Osb.) were characterized for the first time. Further analysis was performed in ‘Cravo’ rangpur lime and ‘Foster’ grapefruit aiming to verify the in vitro morphogenesis of five distinct regions of the epicotyl under different treatments. It was observed the same general pattern of morphogenic gradient along the epicotyl axis in both citrus cultivars, with greater organogenic response as the distance of the explants from the cotyledonary node increased. This morphogenic gradient was influenced by factors related to plant material, composition of the culture medium, and conditions of incubation. The regions of the epicotyl farthest from cotyledons could be used as a source of explants in experiments of genetic transformation of the genotypes evaluated aiming to improve the efficiency of production of transgenic Citrus plants.     

利用计算机软件包鉴别鲜食葡萄品种研究

采用法国葡萄分类学家嘎勒的葡萄表现型分类法中的叶片测量法对南方主栽的12个巨峰系萄萄品种和其他4个常见栽培品种进行了形态学鉴定。通过各品种叶形结构的9个性状参数测定和它们的计算机判别统计分析表明,巨峰系葡萄各品种间的平均判别拟合率为60.95％(26.67％—100％),而巨峰品种与其他非巨峰系品种间的平均判别拟合率可达91.96％(86.60％—100％)。亲缘关系远近也会影响葡萄品种鉴别效率。经过未知品种新样方的鉴别测定,证明所得到的葡萄品种检索表和计算机品种鉴别数据库有助于田间和室内的快速品种鉴别。     

Effect of explants source on shoot proliferation and adventitious regeneration in 10 Buddleia cultivars

Viable shoot cultures and weaned plants were obtained from cultured apical meristems with 10 Buddleia cultivars giving viabilities of 32–72%. The number of shoots produced, the micropropagation rate and the root number produced in vitro was higher in meristem derived shoots compared to those derived from shoot-tips. The subsequent growth rate of meristem derived plants, in the greenhouse, was also greater. The number of roots produced by conventional cuttings collected from meristem derived plants was significantly higher than in cuttings which were collected from plants derived from shoot-tips or from the original stock plants.Endogenous bacteria were not detected in either shoot cultures derived from meristems or in 10-week-old weaned plants derived from meristems whereas those derived from shoot-tips showed the presence of endogenous bacteria when sterilized explants were cultured on nutrient agar or on tryptic soy broth.Factors affecting adventitious bud and shoot production in leaf and internode explants was determined for ‘Lochinch’, ‘Border Beauty’, ‘Ile de France’ and ‘Pink Delight’ using meristem derived shoot cultures. Adventitious shoots appeared after 4 weeks of culture, in both types of explant when cultured on MS supplemented with 0.5–5.0 μM TDZ. The highest percentage regeneration was achieved from bisected internode explants cultured on 0.5 μM TDZ, with 93–100% regeneration among the cultivars whereas BA was less effective. The best response was obtained using 5.0 μM TDZ which gave over 10–11 shoots per explant in all bisected explants for all cultivars.     

Identification and investigation of relationships of mandarin types using isozyme analysis

Sixteen enzyme systems with 19 loci were used to discriminate 19 mandarin cultivars and hybrids. Variability was observed at 12 loci, and all but 3 mandarin types could be discriminated, 2 of which were probably identical. Relatedness within the tangelo and tangor groups was high, probably reflecting their recent origin. Relatedness within the common mandarin group was low, reflecting their multiple origins and long cultivation. Exceptions were cultivars ‘Algerian’ and ‘Beauty of Glen Retreat’, which differed at only 1 locus. Relatedness between the groups was generally low, with the least relatedness between the tangelos and the other groups, probably owing to their grapefruit parent. The ‘Ellendale’ cultivars formed a particularly cohesive group, but with 2 isozyme genotypes, differing at 2 loci, both marketed as ‘Ellendale’. Three of the ‘Ellendale’-type cultivars probably arose by self-pollination of ‘Ellendale’, the rest either by self-pollination or by mutation.     

Genetic diversity and relationships of lotus (Nelumbo) cultivars based on allozyme and ISSR markers

Genetic diversity and genetic relationships of lotus (Nelumbo Adanson) cultivars were evaluated using allozyme and ISSR markers. The samples used covered 11 accessions of possible hybrids between Nelumbo nucifera and Nelumbo lutea and 92 accessions of N. nucifera including 69 flower lotus, 13 rhizome lotus, 5 seed lotus and 5 wild lotus. For allozyme studies, a total of 31 alleles at 23 loci of 18 enzyme systems were detected of which 5 (21.7%) loci Aat, Idh, Mdh-2, Pgd, Sod were polymorphic. The loci of Aat and Idh included two alleles, Mdh-2, Pgd and Sod included three alleles. Eighteen genotypes were detected with the 13 alleles of the 5 polymorphic loci. The parameters of average allele number, observed heterozygosity, expected heterozygosity and Shannon information index of 92 N. nucifera samples were 1.35 ± 0.71, 0.06 ± 0.21, 0.05 ± 0.14, 0.10 ± 023, respectively. Thirteen ISSR primers generated 93 loci, of which 37.63% were polymorphic across all samples. The percentage of polymorphic loci, average allele number, expected heterozygosity and Shannon information index of 92 N. nucifera samples were 26.67%, 1.30 ± 0.46, 0.10 ± 0.18 and 0.15 ± 0.25, respectively for the ISSR data. The ‘Bottleneck effect’ and rapid propagation of clones after the ice ages may explain the low genetic diversity of lotus. The dendrograms based on ISSR and allozymes were not congruent. Based on the ISSR data, the 103 samples were divided into the N. nucifera group (Group I), and the group containing inter-specific hybrids between N. nucifera and N. lutea (Group II). The flower lotus, rhizome lotus, and seed lotus each has multiple sources of origin. Plant size, a criterion commonly used in the classification of cultivars of lotus, is not correlated with genetic variation. Flower color is correlated with the cultivar classification to some degree, but its variation is complex in the hybrids.     

Analysis of the main factors involved in the evaluation of Prunus resistance to Plum pox virus (Sharka) in controlled greenhouse conditions

In countries like Spain or France, quarantine rules force researchers to evaluate the resistance to Sharka (Plum pox virus, PPV) in controlled, isolated conditions. This evaluation method shows important limitations resulting from the management of plants in the controlled conditions, grown in pots with artificial cycles of growth in the greenhouse and cold chamber, alternately. The objective of this study is to analyse different factors that affect the efficiency of the method of evaluation of PPV resistance in controlled greenhouse conditions. The cultivars evaluated as model genotypes were the resistant ‘Stark Early Orange’ and the susceptible ‘Real Fino’ apricot. Furthermore the ‘GF305’ peach was used as a susceptible control. The different studied factors were the inoculation protocol (rootstock or variety inoculation), the grafting success (depending on inoculation method, rootstock–variety combination and date of grafting), and the efficiency of the process in each artificial cycle of growth. Results showed that rootstock inoculation was more effective than inoculation of the variety. As rootstock, the ‘GF305’ seedlings were slightly better than the ‘Real Fino’ seedlings in the inoculation process, but they were quite similar in terms of effectiveness in the evaluation and grafting process. Grafting can be carried out in spring or autumn without having important differences. The global efficiency of the evaluation process was much higher with rootstock inoculation.     

Isoenzyme and microsatellite analysis of Vitis vinifera L. varieties from the Hungarian grape germplasm

The aim of our work was to investigate the genetic diversity of grapevine with biochemical and molecular markers (isoenzyme and SSR). The isoenzyme patterns of 4 enzyme systems (catechol-oxidase, glutamate-oxalacetate-transaminase, acid phosphatase and peroxidase) and the microsatellite profile in 6 loci (VVS2, VVS16, VVMD7, VMC4A1, VMC4G6, VrZag79) of 48 grapevine varieties were analysed. The results with CO, GOT, AcP and PER enzymes were reproducible and the zymograms obtained from the woody stems were independent from the time of sampling during the dormant period of the grape. Based on the isoenzyme patterns of these 4 enzymes most of the investigated varieties (40/48) were identified. A correlation was found between the isoenzyme patterns and the classification to convarietas of the varieties. It was established, that while the varieties of the convarietas pontica differed from those of the convarietas orientalis and occidentalis, the two latter groups could have not been differentiated from each other. Based on the SSR (simple sequence repeat) analyses 46 of the 48 investigated varieties were identified. Even ‘Pinot blanc’ and ‘Pinot gris’ cultivars belonging to the same conculta (Pinot) could be differentiated in their VMC4A1 locus.     

Chill unit models for blackcurrant (Ribes nigrum L.) cultivars ‘Ben Gairn’, ‘Ben Hope’ and ‘Ben Tirran’

Temperate-zone crops require a period of winter chilling to terminate dormancy and ensure adequate bud break the following spring. The exact chilling requirement of blackcurrant (Ribes nigrum), a commercially important crop in northern Europe, is relatively unknown. Chill unit models have been successfully utilized to determine the optimum chilling temperature of a range of crops, with one chill unit equating to 1 h exposure to the optimum temperature for chill satisfaction. Two-year-old R. nigrum plants of the cultivars ‘Ben Gairn’, ‘Ben Hope’ and ‘Ben Tirran’ were exposed to temperatures of −10.1 °C, −3.4 °C, 0.1 °C, 1.5 °C, 2.1 °C, 3.4 °C or 8.9 °C (±0.7 °C) for durations of 0, 2, 4, 6, 8 or 10 weeks and multiple regression analyses used to determine the optimum temperature for chill satisfaction.     

Cross-pollination improves ‘Orri’ mandarin fruit yield

‘Orri’, a selection of ‘Orha’ mandarin [Temple (Citrus temple hort. ex Y. Tanaka) × Dancy (Citrus tangerina hort. ex Tanaka)], is a new high-quality Israeli mandarin which, in the last decade, has become one of the leading varieties in Israel. ‘Orri’ has an excellent taste, the rind is deep orange in color and easily removed, and it contains few or no seeds. However, ‘Orri’ grown in Israel suffers from inadequate yield and no published studies have yet addressed this problem. In the present study we determined that ‘Orri’ productivity depended on conditions being favorable to cross-pollination. Under cross-pollination conditions a positive correlation (R² = 0.97) was found between yield per tree and number of fruits per tree, and more than 90% of the fruits exceeded 60 mm: the most profitable size range. These data suggest that the number of fruits per tree, and not fruit size, is the limiting factor for yield improvement in ‘Orri’ orchards. Studying seed set showed that ‘Michal’ mandarin (Citrus reticulata Blanco) is a compatible pollenizer for ‘Orri’ flowers: the number of seeds per ‘Orri’ fruit increased as the distance from ‘Michal’ trees decreased. The present study demonstrated that cross-pollination of ‘Orri’ resulted in yield improvement, yet at the price of increased seed set.     

Growth responses and ion regulation of four warm season turfgrasses to long-term salinity stress

Increased need for salinity tolerant turfgrasses continues due to increased use of saline water for lawn irrigation and turfgrass establishment on highly saline soil in arid and seashore regions. Turfgrasses growing on saline soil suffer from long-term salinity stress, so this experiment was conducted to study the salinity tolerance, growth, and physiological responses of four warm season turfgrasses [including ‘Diamond’ zoysiagrass (Zoysia matrella (L.) Merr.), ‘Z080’ zoysiagrass (Z. japonica Steud.), ‘C291’ bermudagrass (Cynodon dactylon (L.) Pers), and ‘Adalayd’ seashore paspalum (Paspalum vaginatum Sw.)] to 9 months of salinity stress. Seven salinity levels of irrigation water (0, 90, 180, 360, 540, 720, and 900 mM NaCl) were applied to turfgrasses grown in plastic tubes in a glass room. The salinity tolerance decreased in the following order according to percent green leaf canopy area after 9 months of salinity treatments: ‘Diamond’ > ‘Adalayd’ > ‘C291’ > ‘Z080’. Leaf weight, leaf length, canopy height, shoot density were significantly affected by salinity treatments for all turfgrasses. However, leaf width and/or leaf number per shoot were not affected by salinity in all turfgrasses except ‘Diamond’. Leaf and/or root water contents were also little affected. As salinity increased, leaf and root Na⁺ concentrations and Na⁺/K⁺ rates increased significantly and K⁺ concentrations decreased significantly except that of ‘Adalayd’ leaf. ‘Diamond’ and ‘Z080’ could reduce Na⁺ accumulation in the leaves by salt secretion from salt glands, while ‘Adalayd’ could exclude Na⁺ from the leaves and accumulate K⁺ in the leaves. ‘C291’ exhibited both ion regulation mechanisms, but to much less extent. Different growth responses and ion regulation means of four turfgrasses reflected different salinity tolerance mechanisms.